Effect of the conformational stability of the CH2 domain on the aggregation and peptide cleavage of a humanized IgG

Daisuke Kameoka, Tadashi Ueda, Taiji Imoto

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

To examine the effect of the conformational stability of the CH2 domain on aggregation and peptide cleavage of a humanized IgG1, we carried out size exclusion chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis analyses of incubated sample solutions. By comparing the residual percentage of monomer after incubation at 60 and 80°C at various pH levels, we found that aggregation and peptide cleavage of the humanized IgG1 occurred during long incubation at 60°C under acidic conditions. Next, we confirmed cleavage of the Asp272-Pro273 peptide bond in the CH2 domain. Comparison of the cleavage rates of the IgG1 monomer and a peptide containing the same Asp-Pro sequence revealed that the conformational stability of the CH2 domain retards cleavage of the Asp272-Pro273 peptide bond at 60°C and pH 4.0. The finding of aggregation and peptide cleavage of the humanized IgG1 after long incubation at 60°C under acidic conditions was supported by another finding: there were lower unfolding temperatures of the CH2 domain at pH 4.0 and 5.0. We conclude that the conformational stability of the CH2 domain is closely related to aggregation and peptide cleavage of the humanized IgG1 under acidic conditions. We also found that the 2-[N-morpholino] ethane sulfonate buffer inhibits aggregation of the IgG1 at pH 4.0-5.0 and 7.0-8.0.

Original languageEnglish
Pages (from-to)642-654
Number of pages13
JournalApplied Biochemistry and Biotechnology
Volume164
Issue number5
DOIs
Publication statusPublished - Jul 1 2011

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Peptides
Agglomeration
Immunoglobulin G
aspartyl-proline
Monomers
Morpholinos
Size exclusion chromatography
Sodium dodecyl sulfate
Ethane
Polyacrylates
Electrophoresis
Sodium Dodecyl Sulfate
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Buffers
Gels
Temperature

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology

Cite this

Effect of the conformational stability of the CH2 domain on the aggregation and peptide cleavage of a humanized IgG. / Kameoka, Daisuke; Ueda, Tadashi; Imoto, Taiji.

In: Applied Biochemistry and Biotechnology, Vol. 164, No. 5, 01.07.2011, p. 642-654.

Research output: Contribution to journalArticle

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